Bottom Line:
Comprehensive understanding of the regulation mechanism of Shh in cancer cells is necessary to find an effective approach to selectively block its tumorigenic function.Moreover, in vitro data demonstrated that both NF-κB activation and hypomethylation in promoter region were positively associated with the overexpression of Shh.Furthermore, the biological function data indicated that overexpressed Shh enhanced the self-renewal capacity and migration ability of breast cancer cells, which could be augmented by promoter demethylation and NF-κB activation.

Mentions:
To confirm the correlation among NF-κB nuclear expression, promoter methylation status with Shh expression, several breast cell lines, MCF-7, Bcap37, MDA-MB-436 and MDA-MB-231, were used in our experiments. As shown in Figure2a, MCF-7 cells expressed Shh at a relatively higher level, Bcap37 and MDA-MB-231 cells displayed a moderate expression, but MDA-MB-436 cells expressed Shh at a lower level. Interestingly, a similar pattern was shown in NF-κB nuclear expression, suggesting a positive correlation between NF-κB and Shh. As the transcription factor, NF-κB regulates target gene expression in mRNA level. Thus, we also detected the expression of Shh in mRNA level using quantitative RT-PCR. Consistent with protein expression data, Shh was expressed at a higher level in MCF-7 cells, but not in Bcap37, MDA-MB-436 and MDA-MB-231 cells (see Fig.2b), demonstrating that the crucial regulation of Shh expression is presented in the transcription level.

Mentions:
To confirm the correlation among NF-κB nuclear expression, promoter methylation status with Shh expression, several breast cell lines, MCF-7, Bcap37, MDA-MB-436 and MDA-MB-231, were used in our experiments. As shown in Figure2a, MCF-7 cells expressed Shh at a relatively higher level, Bcap37 and MDA-MB-231 cells displayed a moderate expression, but MDA-MB-436 cells expressed Shh at a lower level. Interestingly, a similar pattern was shown in NF-κB nuclear expression, suggesting a positive correlation between NF-κB and Shh. As the transcription factor, NF-κB regulates target gene expression in mRNA level. Thus, we also detected the expression of Shh in mRNA level using quantitative RT-PCR. Consistent with protein expression data, Shh was expressed at a higher level in MCF-7 cells, but not in Bcap37, MDA-MB-436 and MDA-MB-231 cells (see Fig.2b), demonstrating that the crucial regulation of Shh expression is presented in the transcription level.

Bottom Line:
Comprehensive understanding of the regulation mechanism of Shh in cancer cells is necessary to find an effective approach to selectively block its tumorigenic function.Moreover, in vitro data demonstrated that both NF-κB activation and hypomethylation in promoter region were positively associated with the overexpression of Shh.Furthermore, the biological function data indicated that overexpressed Shh enhanced the self-renewal capacity and migration ability of breast cancer cells, which could be augmented by promoter demethylation and NF-κB activation.